Barbara Ruszkowska-Ciastek

A preliminary evaluation of VEGF-A, VEGFR1 and VEGFR2 in patients with well-controlled type 2 diabetes mellitus

Objective: Decompensated chronic hyperglycemia often leads to late microvascular complications such as retinopathy, diabetic foot syndrome, and diabetic kidney disease. The aim of this study was to determine the concentration of vascular endothelial growth factor A (VEGF-A) and its receptors in patients with well-controlled diabetes. Methods: The study was conducted on 31 patients with well-controlled type 2 diabetes without micro-or macroangiopathy. Thirty healthy volunteers were enrolled in a control group. Serum concentrations of VEGF-A, VEGF receptors 1 and 2 (VEGFR1 and VEGFR2), fasting glucose, and lipid profiles were measured, and the plasma concentration of glycated hemoglobin (HbA1c) was determined. Results: No significant differences were observed between the concentration of VEGF-A, VEGFR1 or VEGFR2 in the subject group and that in the control group. Positive correlations were noted between the levels of VEGF-A, VEGFR2, and triglyceride, and there was a negative correlation between the levels of VEGFR2 and high-density lipoprotein (HDL)-cholesterol in the study group. Conclusions: The concentrations of VEGF-A and its receptors 1 and 2 in patients with well-controlled diabetes are comparable to those of healthy individuals, which may indicate that appropriate control of glucose levels delays the occurrence of vascular complications. A negative correlation between VEGFR2 and HDL-cholesterol levels, and positive correlations between VEGF-A, VEGFR2, and triglyceride levels, suggest that lipid abnormalities occurring in diabetes may be involved in the modulation of angiogenesis.

Effect of uncontrolled hyperglycemia on levels of adhesion molecules in patients with diabetes mellitus type 2.

ObjectiveUncontrolled diabetes has become a major cause of mortality and morbidity by reason of vascular angiopathy. The aim of this study was to evaluate the concentrations of soluble forms of vascular adhesion molecule-1 (sVCAM-1), intercellular adhesion molecule-1 (sICAM-1), E-selectin, and thrombomodulin in patients with well-controlled and uncontrolled diabetes type 2.MethodsThe study was conducted on 62 patients with diabetes. Group I consisted of 35 patients with well-controlled diabetes. The second group included 27 patients with uncontrolled diabetes with micro-albuminuria. A control group was made up of 25 healthy volunteers. The concentrations of sVCAM-1, sICAM-1, sE-selectin, and soluble thrombomodulin were assayed in plasma. Serum concentration of creatinine was measured and the plasma concentrations of fasting glucose and glycated hemoglobin (HbA1c) determined.ResultsLower concentrations of ICAM-1 were found in the group of uncontrolled diabetes patients compared with those with well-controlled disease. In patients with uncontrolled diabetes, VCAM-1 levels were significantly higher compared with the group with well-controlled diabetes. In patients with uncontrolled diabetes a positive correlation was obtained between glomerular filtration rate and sE-selectin and a negative correlation between the levels of creatinine and ICAM-1, although there was a positive correlation between (HbA1c) and ICAM-1.ConclusionThe study confirmed the participation of the inflammatory process associated with impaired vascular endothelial function in the pathogenesis of type 2 diabetes. The opposite effect of uncontrolled hyperglycemia on adhesion molecules suggests different functions of VCAM-1 and ICAM-1 in complications of diabetes.概要目的评估可溶性血管细胞间黏附分子 (sVCAM-1)、可溶性细胞间黏附分子 (sICAM-1)、可溶性选择素E 和可溶性血栓调节蛋白在血糖控制良好和不受控制的2 型糖尿病患者中的水平。创新点对2 型糖尿病患者的血管内皮炎症标记物进行评估。方法62 例糖尿病患者分成两组: 第一组包括35 个血糖控制良好的糖尿病患者, 第二组包括27 个未控制血糖并伴有微蛋白尿的糖尿病患者。对照组由25 名健康志愿者组成。测定血浆中sVCAM-1、sICAM-1、可溶性选择素E 和可溶性血栓调节蛋白的浓度, 同时测定血清肌酐及血浆中空腹血糖和糖化血红蛋白 (HbA1c) 的浓度。结论与血糖控制良好的糖尿病组相比, 未控制血糖组具有相对低的ICAM-1 水平和更高的VCAM-1 水平。未控制血糖组中患者的糖化血红蛋白和ICAM-1 之间呈正相关, 肾小球滤过率和可溶性选择素E 之间呈正相关, 而肌酐和ICAM-1 之间呈负相关。研究证实2 型糖尿病的发病机理中炎症过程的出现与血管内皮功能受损有关。未受控制的高血糖对粘附分子的反向作用表明, 在糖尿病的并发症中VCAM-1和ICAM-1具有不同功能。

Impact of type 2 diabetes on the plasma levels of vascular endothelial growth factor and its soluble receptors type 1 and type 2 in patients with peripheral arterial disease

ObjectiveType 2 diabetes coexistent with lower extremity artery disease (peripheral arterial disease (PAD)) can be observed in numerous patients. The mechanism compensating for ischemia and contributing to healing is angiogenesis—the process of forming new blood vessels. The purpose of this study was to assess the likely impact of type 2 diabetes on the plasma levels of proangiogenic factor (vascular endothelial growth factor A (VEGF-A)) and angiogenesis inhibitors (soluble VEGF receptors type 1 and type 2 (sVEGFR-1 and sVEGFR-2)) in patients with PAD.MethodAmong 46 patients with PAD under pharmacological therapy (non-invasive), we identified, based on medical history, a subgroup with coexistent type 2 diabetes (PAD-DM2+, n=15) and without diabetes (PAD-DM2−, n=31). The control group consisted of 30 healthy subjects. Plasma levels of VEGF-A, sVEGFR-1, and sVEGFR-2 were measured using the enzyme-linked immunosorbent assay (ELISA) method.ResultsThe subgroups of PAD-DM2+ and PAD-DM2−revealed significantly higher concentrations of VEGF-A (P=0.000 007 and P=0.000 000 1, respectively) and significantly lower sVEGFR-2 levels (P=0.02 and P=0.000 01, respectively), when compared with the control group. Patients with PAD and coexistent diabetes tended to have a lower level of VEGF-A and higher levels of sVEGFR-1 and sVEGFR-2 comparable with non-diabetic patients.ConclusionsThe coexistence of type 2 diabetes and PAD is demonstrated by a tendency to a lower plasma level of proangiogenic factor (VEGF-A) and higher levels of angiogenesis inhibitors (sVEGFR-1 and sVEGFR-2) at the same time. Regardless of the coexistence of type 2 diabetes, hypoxia appears to be a crucial factor stimulating the processes of angiogenesis in PAD patients comparable with healthy individuals, whereas hyperglycemia may have a negative impact on angiogenesis in lower limbs.中文概要目 的研究2 型糖尿病对外周动脉疾病患者血浆内的血管内皮生长因子(VEGF-A)及其水溶性受体(sVEGFR-1 和sVEGFR-2)浓度的影响。创新点首次研究了2 型糖尿病对外周动脉疾病患者血浆内sVEGFR-1 和sVEGFR-2 浓度的影响。方 法选取46 个外周动脉疾病患者, 根据有无2 型糖尿病分为糖尿病组(15 例)和无糖尿病组(31 例), 另选30 个健康志愿者为正常对照组。采用酶联免疫吸附法(ELISA)检测他们血浆中VEGF-A及sVEGFR-1 和sVEGFR-2 的浓度, 然后通过对比各组浓度研究2 型糖尿病的影响。结 论与正常对照组相比, 外周动脉疾病患者具有较高的VEGF-A 浓度(2 型糖尿病组 P=0.000 007, 非糖尿病组 P=0.000 000 1)以及较低的sVEGFR-2浓度(2 型糖尿病组 P=0.02, 非糖尿病组 P=0.000 01)。同时, 2 型糖尿病组比非糖尿病组具有较低的VEGF-A 浓度及较高的sVEGFR-1 和sVEGFR-2 浓度。研究结果表明: 无论2 型糖尿病是否共存, 缺氧是导致血管生成的一个关键的刺激因素; 同时, 高血糖状态对下肢的血管生成有抑制作用。

Low-grade risk of hypercoagulable state in patients suffering from diabetes mellitus type 2 *

ObjectiveDiabetes, including type 1 and type 2, is associated with the hypercoagulable state. The aim of this study is to evaluate the concentration of selected hemostatic parameters and vascular endothelial growth factor-A (VEGF-A) in diabetic subjects.MethodsThe study was conducted in 62 patients with diabetes. Group I consisted of 27 patients having uncontrolled diabetes with microalbuminuria and Group II included 35 well-controlled diabetic patients. The control group was made up of 25 healthy volunteers. In the citrate plasma, the concentrations of tissue factor (TF), tissue factor pathway inhibitor (TFPI), thrombin-antithrombin (TAT) complexes, and D-dimer were assayed. Serum concentrations of VEGF-A, lipid profile, creatinine, and plasma fasting glucose were measured and in the versene plasma the concentration of HbA1c was determined.ResultsIn the patients with uncontrolled diabetes, higher concentrations of TF, TFPI, and VEGF-A were observed, as compared with the well-controlled diabetics group and the control group. A significantly lower activity of antiplasmin was reported in patients from Group I as compared with the control group. In Group I, using the multivariate regression analysis, the glomerular filtration rate was independently associated with VEGF-A and dependently associated with total cholesterol.ConclusionsThe study showed higher concentrations of TF and TFPI in the patients with uncontrolled diabetes with microalbuminuria, which is associated with rapid neutralization of the thrombin formation, since TFPI inhibits the complex of TF/VIIa/Ca2+. The manifestation of the above suggestions is the correct TAT complexes and D-dimer, which indicates a low grade of prothrombotic risk in this group of patients, but a higher risk of vascular complications.中文概要目的评估糖尿病病人的止血参数和血管内皮生长因子的浓度。方法62例糖尿病患者分成两组:第一组包括35个血糖控制良好的糖尿病患者,第二组包括27个未控制血糖并伴有微蛋白尿的糖尿病患者。对照组由25名健康志愿者组成。测定血浆中组织因子(TF)、组织因子途径抑制剂(TFPI)、凝血酶抗凝血酶复合物(TAT)和D-二聚体的浓度。同时测定血清中内皮生长因子A(VEGF-A)、血脂、肌酐和血浆空腹血糖及糖化血红蛋白(HbA1c)的浓度。结论研究表明,未控制血糖并伴有微蛋白尿的糖尿病患者具有更高的浓度的TF和TFPI,这与凝血酶形成的快速中和有关。TAT复合物和D-二聚体的正确形成,能保证患者具有一个相对较低级凝血风险,但同时会带来更高的血管并发症的风险。

Circulating endothelial progenitor cells and angiogenic factors in diabetes complicated diabetic foot and without foot complications

INTRODUCTION Data about angiogenic factors in diabetic foot syndrome (DFS) are insufficient. Therefore, in the present study we focus on circulating endothelial progenitor cells (EPCs) and two major angiogenic factors: vascular endothelial growth factor (VEGF-A) and fibroblast growth factor (FGF-2) in patients with DFS. MATERIALS AND METHODS We included 75 subjects: 45 patients with type 2 diabetes and 30 controls. The study group was divided into 2 subgroups: 23 patients with diabetic foot and 22 patients without diabetic complications. The concentration of VEGF-A, soluble VEGF receptor 2 (sVEGF-R2) and FGF-2 were measured in plasma samples. The number of circulating EPCs was determined in peripheral venous blood. The number of endothelial progenitor cells was measured with FACSCalibur flow cytometer using monoclonal antibodies directed against antigens specific for EPCs. RESULTS In our study we observed significant higher levels of VEGF-A and FGF-2 and lower sVEGF-R2 concentration in patients with T2DM compared to healthy subjects. The conducted analysis showed decreased levels of VEGF-A and elevated levels of FGF-2 in patients with DM complicated DFS compared to diabetic patients without DFS. Increased circulating EPCs number was reported in patients with DFS, and the difference was almost statistically significant. CONCLUSIONS The high concentration of VEGF-A and FGF-2, and a positive correlation between them indicate their participation in the process of angiogenesis in T2DM. Decreased sVEGF-R2 may result from inactivation of VEGF-A during complexes formation.

CRP, but not TNF-α or IL-6, decreases after weight loss in patients with morbid obesity exposed to intensive weight reduction and balneological treatment.

ObjectiveThe aim of this study was to evaluate the concentrations of C-reactive protein (CRP), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and the degree of homeostasis model assessment-insulin resistance (HOMA-IR) in patients with morbid obesity exposed to a three-week low-calorie diet and balneotherapy.MethodsThe study included 33 patients (25 females and 8 males; mean age 46 years) with body mass index (BMI) values of >40 kg/m2. Evaluations of CRP, IL-6, TNF-α, lipid profile, HOMA-IR, and fasting glucose were carried out before (baseline data) and three weeks after the treatment. The control group consisted of 20 healthy volunteers (15 females and 5 males) with a mean age of 39 years and BMI values of ≤24.9 kg/m2.ResultsIn the blood of patients with morbid obesity we found significantly elevated levels of CRP, TNF-α, triglycerides, HOMA-IR and fasting glucose, but a decreased level of high density lipoprotein (HDL)-cholesterol, compared with the healthy individuals. The treatment resulted in about a 9.4% reduction in body weight from 122.5 to 111.0 kg and a significant decrease in the concentration of CRP, but no change in TNF-α or IL-6. HOMA-IR was significantly reduced.ConclusionThe decrease in CRP level without changes in TNF-α or IL-6 concentrations after the low-calorie diet and balneological treatment, suggests that an essential amount of adipose tissue must be removed before proper adipocyte function is restored. The decrease in HOMA-IR indicates an improvement in insulin sensitivity, which is beneficial in obese patients.概要目的评估病态肥胖患者在低热量饮食和浸浴治疗前后, C 反应蛋白 (CRP)、肿瘤坏死因子α (TNF-α)、白细胞介素-6 ( IL-6) 以及胰岛素抵抗指数 (HOMA-IR) 的水平变化。创新点研究炎症反应对病态肥胖的影响。方法本研究包括33 个病例和20 个正常对照, 对治疗前后的CRP、IL-6、TNF-α、血脂谱、HOMA-IR以及空腹血糖水平进行评估。结论与对照组相比, 低热量饮食和浸浴治疗后, 病态肥胖患者的CRP水平显著降低, 而TNF-α 和IL-6的水平却没有显著变化, 这表明脂肪细胞的正常功能恢复之前必须先去除一定量的脂肪组织。HOMA-IR 水平的降低说明对胰岛素的敏感性有所提高, 这有利于肥胖患者的治疗。

Selected parameters of hemostasis in patients with myeloproliferative neoplasms.

Hemostatic disorders are a major clinical problem in patients with myeloproliferative neoplasms (MPNs) and they are the second most common cause of death in MPN patients, after infections. The aim of this study was to assess the fibrinolytic potential of the blood of patients with MPNs. The study involved 112 patients with MPNs diagnosed at the Hematology Clinic Dr J. Biziel University Hospital No. 2 in Bydgoszcz, Poland. The study group included 63 patients with essential thrombocythemia, 29 with polycythemia vera, 11 with chronic myelogenous leukemia (CML) and nine with primary myelofibrosis. The control group consisted of 25 healthy volunteers who were age and sex-matched. The following parameters were determined: concentration of tissue plasminogen activator antigen (t-PA:Ag), plasminogen activator inhibitor type 1 antigen concentration (PAI-1:Ag), D-dimer, thrombin–antithrombin complexes, fibrinogen, activated partial thromboplastin time and international normalized ratio. The study showed significantly increased t-PA:Ag, PAI-1:Ag and D-dimer levels in patients with MPNs. Moreover, we found increased concentrations of thrombin–antithrombin complexes and fibrinogen, as well as elevated platelet counts. Detailed analysis revealed that t-PA:Ag concentration was elevated in patients with essential thrombocythemia, CML and polycythemia vera. Concentration of PAI-1:Ag was increased in patients with essential thrombocythemia and polycythemia vera; D-dimer was significantly higher in essential thrombocythemia, polycythemia vera, CML and primary myelofibrosis patients. Increased concentrations of t-PA:Ag and D-dimer indicate secondary activation of the fibrinolytic system in patients with MPNs. Elevated levels of PAI-1 in MPN patients may result from its increased production by elevated number of activated platelets and vascular endothelial damage. PAI-1 by having an inhibitory effect on fibrinolysis manifests its procoagulant activity.

Heterogeneity of cardiovascular risk factors profile in non-diabetic women 2–24 months post gestational diabetes mellitus.

Abstract Previously gestational diabetic (pGDM) women are characterized by high cardiovascular risk (CVR). The aim of this study was to assess the CVR markers levels in non-diabetic pGDM women in relation to time postpartum and to soluble E-selectin (sES) level. We investigated 125 women aged 18–40 years with a history of GDM between 2 and 24 months after their pregnancy. We evaluated age, body mass index (BMI), waist circumference, glucose levels during the oral glucose tolerance test (OGTT), levels of insulin and the parameters of endothelial dysfunction, fibrinolysis activity, low-grade systemic inflammation and lipid profiles. Prediabetes was identified in 38 women (30%), while in the remaining women OGTT results were normal. The tests performed >6 months revealed decreased hs-CRP (p = 0.01), sICAM-1 (p = 0.01), and elevated sES (p = 0.01) >12 months after adjustment for age, BMI, waist circumference and 2 h OGTT glucose. In the subgroup tested ≤12 months after an index pregnancy sES was independently associated with hs-CRP (p < 0.0001) and triglycerides (p = 0.0139). No association was found between sES and remaining parameters in women tested >12 months postpartum. We conclude that the period 2–24 months post GDM is heterogeneous with respect to the CVR markers. The plasma level of hs-CRP could be useful as an important cardiovascular risk marker up to 12 months postpartum in non-diabetic pGDM women.

Gestational Diabetes Mellitus Worsens the Profile of Cardiometabolic Risk Markers and Decrease Indexes of Beta-Cell Function Independently of Insulin Resistance in Nondiabetic Women with a Parental History of Type 2 Diabetes

Background. Women with a history of both parental type 2 diabetes (pt2DM) and previous gestational diabetes (pGDM) represent a group at high risk of cardiovascular events. We hypothesized that pGDM changes cardiometabolic risk markers levels as well as theirs associations with glucose indices in nondiabetic pt2DM women. Methods. Anthropometric parameters, glucose regulation (OGTT), insulin resistance (HOMA-IR), beta-cell function, lipid levels, parameters of endothelial dysfunction, and inflammation were evaluated in 55 women with pt2DM, 40 with both pt2DM and pGDM 2–24 months postpartum, and 35 controls. Results. Prediabetes was diagnosed more frequently in women with both pt2DM and pGDM in comparison with women with only pt2DM (10 versus 8, P = 0.04). The pGDM group had higher LDL-cholesterol, sICAM-1, tPa Ag, fibrinogen, and lower beta-cell function after adjustment for HOMA-IR, in comparison with pt2DM group. In pt2DM group postchallenge glucose correlated independently with hsCRP and in pGDM group fasting glucose with HOMA-IR. Conclusions. pGDM exerts a combined effect on cardiometabolic risk markers in women with pt2DM. In these women higher LDL-cholesterol, fibrinogen, sICAM-1, tPa Ag levels and decreased beta cell function are associated with pGDM independently of HOMA-IR index value. Fasting glucose is an important cardiometabolic risk marker and is independently associated with HOMA-IR.

Effect of a 3-Week Low-Calorie Diet and Balneological Treatment on Selected Coagulation Parameters in Morbidly Obese Patients

BACKGROUND The most common cause of death among people with obesity are cardiovascular complications as a result of a hypercoagulability state. OBJECTIVES The purpose of the study was to assess the potential of coagulation system activation depending on the tissue factor and to analyze of the influence of a 3-week low-calorie diet and balneological treatment on selected coagulation parameters in morbidly obese patients. MATERIAL AND METHODS The study included 36 patients (28 females and 8 males; mean age 46) with the value of BMI > 40 kg/m2. The study was designed in two stages: baseline and after 21-days. The evaluation of tissue factor (TF), tissue factor pathway inhibitor (TFPI), von Willebrand factor (vWF), fibrinogen, D-dimer, thrombin-antithrombin complexes (TAT), and the activity of antithrombin (AT) was performed in patients before and after the treatment. The control group consisted of 24 healthy volunteers (16 females and 8 males) at a mean age of 39 with BMI . 24.9 kg/m2. RESULTS There were significantly higher levels of TF, TFPI, fibrinogen, TAT complexes and D-dimer in the study group as compared to the controls. Moreover, there were no significant changes in the parameters studied before and after the treatment. In the group of obese patients, there were significant positive correlations between the concentrations of vWF and BMI and BMI changes and a significant negative correlation between the WHR changes and TFPI concentration. CONCLUSIONS The study confirmed that morbidly obese patients represent a high risk of hypercoagulability state, despite no clinical evidence, which could be due to the great inhibitory potential of TFPI in suppressing the extrinsic pathway of the coagulation system. However, the lack of effect of the 3-week exposure to the LCD and balneological treatment in morbidly obese subjects indicates that substantial fat mass must be reduced before adequate hemostasis is re-established.